Sugano Yasushi, Matsushima Yuko, Tsuchiya Katsunori, Aoki Hirokazu, Hirai Mitsuyo, Shoda Makoto
R1-30 Chemical Resources Laboratory, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama, 226-8503, Japan.
Biodegradation. 2009 Jun;20(3):433-40. doi: 10.1007/s10532-008-9234-y. Epub 2008 Nov 15.
The reactants produced by action of a purified unique dye-decolorizing peroxidase, DyP, on a commercial anthraquinone dye, Reactive Blue 5, were investigated using electrospray ionization mass spectrometry (ESI-MS), thin-layer chromatography (TLC), and (1)H- and (13)C- nuclear magnetic resonance (NMR). The results of ESI-MS analysis showed that phthalic acid, a Product 2 (molecular weight 472.5), and a Product 3 (molecular weight 301.5), were produced. Product 2 and Product 3 were generated by usual peroxidase reaction, whereas phthalic acid was generated by hydrolase- or oxygenase-catalyzed reaction. One potential associated product, o-aminobenzene sulfonic acid, was found to be converted to 2,2'-disulfonyl azobenzene by ESI-MS and NMR analyses. From these results, we propose, for the first time, the degradation pathway of an anthraquinone dye by the enzyme DyP.
利用电喷雾电离质谱（ESI-MS）、薄层色谱（TLC）以及氢-1和碳-13核磁共振（NMR），对纯化后的独特染料脱色过氧化物酶（DyP）作用于商业蒽醌染料活性蓝5所产生的反应物进行了研究。ESI-MS分析结果表明，生成了邻苯二甲酸、产物2（分子量472.5）和产物3（分子量301.5）。产物2和产物3是通过常见的过氧化物酶反应生成的，而邻苯二甲酸是通过水解酶或加氧酶催化反应生成的。通过ESI-MS和NMR分析发现，一种潜在的相关产物邻氨基苯磺酸被转化为2,2'-二磺酰基偶氮苯。基于这些结果，我们首次提出了酶DyP对蒽醌染料的降解途径。
